Investigation of rhabdomyosarcoma cell-cell electrofusion

Cell-cell fusion is an important natural and engineered process for in-depth studies into hybridomas, developmental biology, immunology and various cellular therapies. It is also a powerful tool for analysis of gene expression, chromosomal mapping, antibody production, cloning mammals and cancer immunotherapy. However, research so far has primarily focused on cell models such as C.elegans, drosophila, myoblasts, spleen-myeloma cell hybrids and various plant protoplasts. Rhabdomyosarcoma cells are a rare form of musculoskeletal cancer cells found in the head/neck, and other less skeletal areas of the human cancer patient’s body. These cells do not normally undergo fusion naturally. Therefore, if these cells can be engineered to fuse, the technique is expected to be effective for other cell types. Among all the techniques of cell fusion, electroporation was chosen due to its high versatility, yield and viability. By coupling the cells with this technique, the effects on cell proliferation, growth pattern and hybridoma count were investigated. Overall, the experimental results showed that an adequate electrical stimulation successfully helped to facilitate the fusion and proliferation of the cells, and the DC current produced the highest fusion yield and proliferation. Under DC current, a fusion yield of 27% and 38% was obtained, compared to 4% and 13% with no electrical current respectively. The work done proved that high yield and proliferation rate could be achieved using a design with two parallel thin gold electrodes sputtered onto the flask, making this an efficient and effective cellular engineering system.MASTER OF ENGINEERING (MAE